Proteomic Profiles of Exosomes of Septic Patients Presenting to the Emergency Department Compared to Healthy Controls
Abstract
:1. Introduction
2. Experimental Section
2.1. Patient Recruitment Criteria
2.2. Isolation and Characterization of Plasma Exosomes
2.3. Proteomics
2.4. Statistical Methods
2.5. Pathway Analysis
3. Results
3.1. Patients and Controls
3.2. Characterization of Exosomes
3.3. Proteomics: Comparison of Control vs. Sepsis Patients: Batch Effect Correction
3.4. Differential Expression Analysis of Sepsis vs. Controls
3.5. Enrichment Analysis of Proteomic Data
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Data Availability
References
- Lydon, E.C.; Ko, E.R.; Tsalik, E.L. The host response as a tool for infectious disease diagnosis and management. Expert Rev. Mol. Diagn. 2018, 18, 723–738. [Google Scholar] [CrossRef] [PubMed]
- Poore, G.D.; Ko, E.R.; Valente, A.; Henao, R.; Sumner, K.; Hong, C.; Burke, T.W.; Nichols, M.; McClain, M.T.; Huang, E.S.; et al. A miRNA Host Response Signature Accurately Discriminates Acute Respiratory Infection Etiologies. Front. Microbiol. 2018, 9, 2957. [Google Scholar] [CrossRef] [PubMed]
- Yang, W.E.; Suchindran, S.; Nicholson, B.P.; McClain, M.T.; Burke, T.W.; Ginsburg, G.S.; Harro, C.D.; Chakraborty, S.; Sack, D.A.; Woods, C.W.; et al. Transcriptomic Analysis of the Host Response and Innate Resilience to EnterotoxigenicEscherichia coliInfection in Humans. J. Infect. Dis. 2016, 213, 1495–1504. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Paoli, C.J.; Reynolds, M.A.; Sinha, M.; Gitlin, M.; Crouser, E. Epidemiology and Costs of Sepsis in the United States—An Analysis Based on Timing of Diagnosis and Severity Level. Crit. Care Med. 2018, 46, 1889–1897. [Google Scholar] [CrossRef]
- Park, E.J.; Appiah, M.G.; Myint, P.K.; Gaowa, A.; Kawamoto, E.; Shimaoka, M. Exosomes in Sepsis and Inflammatory Tissue Injury. Curr. Pharm. Des. 2020, 25, 4486–4495. [Google Scholar] [CrossRef]
- Xin, H.; Li, Y.; Chopp, M. Exosomes/miRNAs as mediating cell-based therapy of stroke. Front. Cell Neurosci. 2014, 8, 377. [Google Scholar] [CrossRef] [Green Version]
- Im, Y.; Yoo, H.; Lee, J.Y.; Park, J.; Suh, G.Y.; Jeon, K. Association of plasma exosomes with severity of organ failure and mortality in patients with sepsis. J. Cell Mol. Med. 2020, 24, 9439–9445. [Google Scholar] [CrossRef]
- Azevedo, L.C.P.; Janiszewski, M.; Pontieri, V.; Pedro, M.D.A.; Bassi, E.; Tucci, P.J.F.; Francisco, Y. Platelet-derived exosomes from septic shock patients induce myocardial dysfunction. Crit. Care 2007, 11, R120. [Google Scholar] [CrossRef] [Green Version]
- Azevedo, L.C.; Pedro, M.D.A.; Souza, L.C.; De Souza, H.P.; Janiszewski, M.; Da Luz, P.L.; Francisco, Y. Oxidative stress as a signaling mechanism of the vascular response to injury The redox hypothesis of restenosis. Cardiovasc. Res. 2000, 47, 436–445. [Google Scholar] [CrossRef] [Green Version]
- Gambim, M.H.; Carmo, A.D.O.D.; Marti, L.C.; Veríssimo-Filho, S.; Lopes, L.R.; Janiszewski, M. Platelet-derived exosomes induce endothelial cell apoptosis through peroxynitrite generation: Experimental evidence for a novel mechanism of septic vascular dysfunction. Crit. Care 2007, 11, R107. [Google Scholar] [CrossRef] [Green Version]
- Janiszewski, M.; Carmo, A.O.D.; Pedro, M.A.; Silva, E.; Knobel, E.; Laurindo, F.R.M. Platelet-derived exosomes of septic individuals possess proapoptotic NAD(P)H oxidase activity: A novel vascular redox pathway. Crit. Care Med. 2004, 32, 818–825. [Google Scholar] [CrossRef] [PubMed]
- Real, J.M.; Ferreira, L.R.P.; Esteves, G.H.; Koyama, F.C.; Dias, M.V.S.; Bezerra-Neto, J.E.; Cunha-Neto, E.; Machado, F.R.; Salomão, R.; Azevedo, L.C.P. Exosomes from patients with septic shock convey miRNAs related to inflammation and cell cycle regulation: New signaling pathways in sepsis? Crit. Care 2018, 22, 68. [Google Scholar] [CrossRef] [Green Version]
- Gao, K.; Jin, J.; Huang, C.; Li, J.; Luo, H.; Li, L.; Huang, Y.; Jiang, Y. Exosomes Derived From Septic Mouse Serum Modulate Immune Responses via Exosome-Associated Cytokines. Front. Immunol. 2019, 10, 1560. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Xu, Y.; Ku, X.; Wu, C.; Cai, C.; Tang, J.; Yan, W. Exosomal proteome analysis of human plasma to monitor sepsis progression. Biochem. Biophys. Res. Commun. 2018, 499, 856–861. [Google Scholar] [CrossRef]
- Leisman, D.; Doerfler, M.E.; Ward, M.F.; Masick, K.D.; Wie, B.; Gribben, J.L.; Hamilton, E.; Klein, Z.; Bianculli, A.R.; Akerman, M.B.; et al. Survival Benefit and Cost Savings From Compliance With a Simplified 3-Hour Sepsis Bundle in a Series of Prospective, Multisite, Observational Cohorts. Crit. Care Med. 2017, 45, 395–406. [Google Scholar] [CrossRef]
- Keller, A.; Nesvizhskii, A.I.; Kolker, A.E.; Aebersold, R. Empirical Statistical Model To Estimate the Accuracy of Peptide Identifications Made by MS/MS and Database Search. Anal. Chem. 2002, 74, 5383–5392. [Google Scholar] [CrossRef] [PubMed]
- Nesvizhskii, A.I.; Keller, A.; Kolker, E.; Aebersold, R. A Statistical Model for Identifying Proteins by Tandem Mass Spectrometry. Anal. Chem. 2003, 75, 4646–4658. [Google Scholar] [CrossRef]
- Johnson, W.E.; Li, C.; Rabinovic, A. Adjusting batch effects in microarray expression data using empirical Bayes methods. Biostatistics 2006, 8, 118–127. [Google Scholar] [CrossRef]
- Ritchie, M.E.; Phipson, B.; Wu, D.; Hu, Y.; Law, C.W.; Shi, W.; Smyth, G.K. limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res. 2015, 43, e47. [Google Scholar] [CrossRef]
- Hochberg, Y.; Benjamini, Y. More powerful procedures for multiple significance testing. Stat. Med. 1990, 9, 811–818. [Google Scholar] [CrossRef]
- Krämer, A.; Green, J.; Pollard, J.; Tugendreich, S. Causal analysis approaches in Ingenuity Pathway Analysis. Bioinformatics 2013, 30, 523–530. [Google Scholar] [CrossRef]
- Ingenuity Pathway Analysis (IPA). Available online: https//www.qiagenbioinformatics.com/products/ingenuity-pathway-analysis (accessed on 1 April 2020).
- Knaus, W.A.; Draper, E.A.; Wagner, D.P.; Zimmerman, J.E. Apache ii: A severity of disease classification system. Crit. Care Med. 1985, 13, 818–829. [Google Scholar] [CrossRef]
- Vincent, J.L.; Moreno, R.; Takala, J.; Willatts, S.; De Mendonca, A.; Bruining, H.; Reinhart, C.K.; Suter, P.M.; Thijs, L.G. The sofa (sepsis-related organ failure assessment) score to describe organ dysfunction/failure. On behalf of the working group on sepsis-related problems of the european society of intensive care medicine. Intensive Care Med. 1996, 22, 707–710. [Google Scholar] [CrossRef]
- Annane, D.; Renault, A.; Brun-Buisson, C.; Mégarbane, B.; Quenot, J.-P.; Siami, S.; Cariou, A.; Forceville, X.; Schwebel, C.; Martin-Loeches, I.; et al. Hydrocortisone plus Fludrocortisone for Adults with Septic Shock. N. Engl. J. Med. 2018, 378, 809–818. [Google Scholar] [CrossRef] [PubMed]
- Nguyen, H.T.; Jaehne, A.K.; Jayaprakash, N.; Semler, M.W.; Hegab, S.; Yataco, A.O.C.; Tatem, G.; Salem, D.; Moore, S.; Boka, K.; et al. Early goal-directed therapy in severe sepsis and septic shock: Insights and comparisons to ProCESS, ProMISe, and ARISE. Crit. Care 2016, 20, 160. [Google Scholar] [CrossRef] [Green Version]
- Peake, S.L.; Bailey, M.; Bellomo, R.; Cameron, P.A.; Cross, A.; Delaney, A.; Finfer, S.; Higgins, A.; Jones, D.A.; Myburgh, J.A.; et al. Australasian resuscitation of sepsis evaluation (arise): A multi-centre, prospective, inception cohort study. Resuscitation 2009, 80, 811–818. [Google Scholar] [CrossRef] [PubMed]
- Latten, G.; Claassen, L.; Jonk, M.; Cals, J.W.L.; Muris, J.W.M.; Stassen, P.M. Characteristics of the prehospital phase of adult emergency department patients with an infection: A prospective pilot study. PLoS ONE 2019, 14, e0212181. [Google Scholar] [CrossRef] [PubMed]
- Nguyen, H.B.; Loomba, M.; Yang, J.J.; Jacobsen, G.; Shah, K.; Otero, R.M.; Suarez, A.; Parekh, H.; Jaehne, A.K.; Rivers, E.P. Early lactate clearance is associated with biomarkers of inflammation, coagulation, apoptosis, organ dysfunction and mortality in severe sepsis and septic shock. J. Inflamm. 2010, 7, 6. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Rivers, E.P.; Jaehne, A.K.; Nguyen, H.B.; Papamatheakis, D.G.; Singer, D.; Yang, J.J.; Brown, S.; Klausner, H. Early biomarker activity in severe sepsis and septic shock and a contemporary review of immunotherapy trials: Not a time to give up, but to give it earlier. Shock 2013, 39, 127–137. [Google Scholar] [CrossRef]
- Nguyen, H.B.; Rivers, E.P.; Knoblich, B.P.; Jacobsen, G.; Muzzin, A.; Ressler, J.A.; Tomlanovich, M.C. Early lactate clearance is associated with improved outcome in severe sepsis and septic shock. Crit. Care Med. 2004, 32, 1637–1642. [Google Scholar] [CrossRef]
- Sander, L.E.; Sackett, S.D.; Dierssen, U.; Beraza, N.; Linke, R.P.; Muller, M.; Blander, J.M.; Tacke, F.; Trautwein, C. Hepatic acute-phase proteins control innate immune responses during infection by promoting myeloid-derived suppressor cell function. J. Exp. Med. 2010, 207, 1453–1464. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Gabay, C.; Kushner, I. Acute-phase proteins and other systemic responses to inflammation. N. Engl. J. Med. 1999, 340, 448–454. [Google Scholar] [CrossRef] [PubMed]
- Sproston, N.R.; Ashworth, J.J. Role of c-reactive protein at sites of inflammation and infection. Front. Immunol. 2018, 9, 754. [Google Scholar] [CrossRef] [PubMed]
- Volanakis, J.E. Human c-reactive protein: Expression, structure, and function. Mol. Immunol. 2001, 38, 189–197. [Google Scholar] [CrossRef]
- Torzewski, M.; Rist, C.; Mortensen, R.F.; Zwaka, T.P.; Bienek, M.; Waltenberger, J.; Koenig, W.; Schmitz, G.; Hombach, V.; Torzewski, J. C-reactive protein in the arterial intima: Role of C-reactive protein receptor-dependent monocyte recruitment in atherogenesis. Arter. Thromb. Vasc. Boil. 2000, 20, 2094–2099. [Google Scholar] [CrossRef] [Green Version]
- Urieli-Shoval, S.; Linke, R.P.; Matzner, Y. Expression and function of serum amyloid A, a major acute-phase protein, in normal and disease states. Curr. Opin. Hematol. 2000, 7, 64–69. [Google Scholar] [CrossRef]
- Sjaastad, F.V.; Condotta, S.A.; Kotov, J.A.; Pape, K.A.; Dail, C.; Danahy, D.B.; Kucaba, T.A.; Tygrett, L.T.; Murphy, K.A.; Cabrera-Perez, J.; et al. Polymicrobial Sepsis Chronic Immunoparalysis Is Defined by Diminished Ag-Specific T Cell-Dependent B Cell Responses. Front. Immunol. 2018, 9, 2532. [Google Scholar] [CrossRef]
- Vassilev, T.; Bauer, M. Passive immunotherapy of sepsis with intravenous immune globulin: Not all IVIg preparations are created equal. Crit. Care 2012, 16, 407. [Google Scholar] [CrossRef] [Green Version]
- Werdan, K.; Pilz, G. Supplemental immune globulins in sepsis: A critical appraisal. Clin. Exp. Immunol. 1996, 104, 83–90. [Google Scholar] [CrossRef]
Batch 1 | Batch 2 | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
Patient 1 | Patient 2 | Patient 3 | Control 1 | Control 2 | Control 3 | Patient ATB1 | Patient ATB2 | Patient ATB24 | Patient ATB46 | Control 1 | Control 2 | |
Gender | Male | Female | Female | Male | Female | Female | Male | Male | Male | Male | Male | Male |
Age | 61 | 74 | 52 | 55 | 24 | 39 | 64 | 81 | 68 | 41 | 60 | 54 |
Race | African American | African American | Caucasian | Caucasian | Asian | Caucasian | African American | African American | African American | Unknown | African American | African American |
Primary Source | Blood | Lung, Free Intraabdominal air | Urogenital | None | None | None | Lung | Abdomen | Lung, peritoneal dialysis cath | Blood | None | None |
Sepsis Class | Shock | Shock | Severe | Healthy | Healthy | Healthy | Severe | Severe | Shock | Shock | Healthy | Healthy |
30 Day Outcome | Dead | Alive | Alive | Alive | Alive | Alive | Alive | Alive | Alive | Dead | Alive | Alive |
Length of Stay | 17 | 7 | 4 | 6 | 6 | 28 | 11 | |||||
Day 1 SOFA Score | 13 | 9 | 14 | 4 | 4 | 12 | 10 | |||||
Day1 APACHE Score | 24 | 33 | 15 | 11 | 30 | 34 | 27 | |||||
Vasopressor Use | Yes | Yes | No | No | Ni | Yes | Yes | |||||
Culture Positive | Wound | Urine, Blood | Urine | Nasal Swab | Urine | None | None | |||||
Organism | Pseudomonas aeruginosa | Candida albicans, Staph Coag (-) | Escherichia Coli | Influenza A | Enterococcus Sp. | None | None | |||||
Highest Lactate mg/ dL | 10.8 | 10.0 | 4.5 | 8.5 | 1.1 | 3.1 | 1.6 |
© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Morris, D.C.; Jaehne, A.K.; Chopp, M.; Zhang, Z.; Poisson, L.; Chen, Y.; Datta, I.; Rivers, E.P. Proteomic Profiles of Exosomes of Septic Patients Presenting to the Emergency Department Compared to Healthy Controls. J. Clin. Med. 2020, 9, 2930. https://0-doi-org.brum.beds.ac.uk/10.3390/jcm9092930
Morris DC, Jaehne AK, Chopp M, Zhang Z, Poisson L, Chen Y, Datta I, Rivers EP. Proteomic Profiles of Exosomes of Septic Patients Presenting to the Emergency Department Compared to Healthy Controls. Journal of Clinical Medicine. 2020; 9(9):2930. https://0-doi-org.brum.beds.ac.uk/10.3390/jcm9092930
Chicago/Turabian StyleMorris, Daniel C., Anja K. Jaehne, Michael Chopp, Zhanggang Zhang, Laila Poisson, Yalei Chen, Indrani Datta, and Emanuel P. Rivers. 2020. "Proteomic Profiles of Exosomes of Septic Patients Presenting to the Emergency Department Compared to Healthy Controls" Journal of Clinical Medicine 9, no. 9: 2930. https://0-doi-org.brum.beds.ac.uk/10.3390/jcm9092930